Quantification of Axin and β-Catenin Dynamics

Images of cells were taken on confocal microscopes, a Zeiss 780 confocal microscope or a 3i spinning disk confocal microscope (CSU-W1 SoRa). 40 × objective lens were used. For quantification, areas on each slide contain at least 150 transfected cells and 1000 non-transfected cells were imaged using the 3i spinning disk confocal microscope. Quantification of cell intensity and puncta size were done with CellProfiler (version 3.1.8)^{46 (link)}. Due to limit of resolution, only puncta with a diameter equal or greater than 0.5 µm were identified and their size measured. mCherry-Axin intensity and β-catenin intensity were normalized to the endogenous β-catenin intensity in non-transfected cells (average intensity of over 1000 cells) on each slide before comparing across slides. Statistics and Plots were done with GraphPad Prism. The data did not pass a normal distribution test. Thus non-parametric one-way ANOVA (Kruskal–Wallis one-way analysis) test and Dunn’s multiple comparisons with correction were used.